The invention pertains to a method for stabilising a human blood protein or human blood plasma protein with a molecular weight of >10 KDa, a composition in solid or liquid state of human blood protein or human blood plasma protein with a molecular weight of >10 KDa and the use of melezitose for stabilisation of a human blood protein or human blood plasma protein.
The stabilisation of therapeutic proteins is a major challenge for the formulation scientists in the pharmaceutical industry today. There are many kinds of stresses that can cause both reversible and irreversible changes to the proteins, such as aggregation, precipitation or denaturation. These difficulties call for the need of agents that stabilize these delicate proteins. Formulation development is a critical step, requiring careful selection of excipients to provide a high yield of protein activity during the purification process as well as during the pharmaceutical process and as a final product. In particular this is true for human blood proteins and human blood plasma proteins.
One of the most widely used stabilizers for protein formulations are carbohydrates, also called saccharides. Carbohydrates are built of linked basic carbohydrate components called monosaccharides, and can be of different length and can thus have different characteristics.
Sucrose and trehalose, the two most commonly used stabilizers, are both disaccharides, hence composed of two monosaccharides.
As compared to two of the most commonly used carbohydrate stabilizers sucrose and trehalose, which are disaccharides, melezitose is a trisaccharide. It is generally indicated [Wang W, Lyophilisation and development of solid protein pharmaceuticals, Int J Pharm 203, 1-60, 2000; Carpenter J. F., Chang B. S., Garzon-Rodriguez W., Randolph T. W., Rational design of stable lyophilized protein formulations: Theroy and practice, chapter 5, ed Carpenter and Manning, Kluiwer Academic/Plenum Publishers, New York, 2002] that disaccharides are the first choice for stabilisation of proteins both in solution and in lyophilized state. Some disaccharides, such as lactose or maltose, are reducing sugars that can degrade proteins via the Malliard reaction during storage in the solid state. If larger saccharides are used as stabilizers in lyophilized preparations, literature suggests that these are less efficient due to steric hinderance of the protein-stabilizer interaction [Carpenter J. F., Chang B. S., Garzon-Rodriguez W., Randolph T. W., Rational design of stable lyophilized protein formulations: Theroy and practice, chapter 5, ed Carpenter and Manning, Kluiwer Academic/Plenum Publishers, New York, 2002].
The review article Wang, W., International Journal of Pharmaceutics, 203 (2000) 1-60, “Lyophilization and development of solid protein pharmaceuticals”, discloses i.a. that maltose, glucose and maltotriose could increase the recovery of catalase activity at 1 mg ml−1, but maltopentaose, maltohexaose, and maltoheptaose were not as effective. The ineffectiveness of larger saccharides suggests that protein stabilization by sugars may depend on the glucoside side chain length of the sugar that may interfere with intermolecular hydrogen-bonding between stabilizing sugars and proteins. This review article recommends disaccharides as stabilizers (p. 9/10).
WO-A-2003/086443 discloses the use of carbohydrates including stachyose, melezitose, and various mono- and disaccharides for preparation of intranasally administerable polypeptide preparations. The sugars serve as agents to reduce the effects of shear stress during spraying.
WO-A-86/04486 discloses chromatographic purification of i. a. factor VIII wherein melezitose is used as a hydration additive during the chromatographic process.
WO-A-91/18091 discloses a method of preserving delicate biological substances or organic compounds (a) in a dry state and/or (b) at elevated temperatures and/or (c) under irradiation comprises incorporating in a system containing the said substances or compounds a sugar or a sugar derivative selected from (i) a non reducing glycoside of a polyhydroxy compounds selected from sugar alcohols and other straight chain polyalcohols, or (ii) a non-reducing oligosaccharide selected from raffinose, stachyose and melezitose.
Mollmann, S. H. et al reports in Drug Dev. Ind. Pharm. 2006 July; (6):765-78 about the stability of insulin in solid formulations containing melezitose and starch.